Objective: To identify the antigenic epitope regions recognized by anti-synuclein-gamma monoclonal antibodies.
Methods: Six expression vectors of truncated SNCG were constructed and the truncated SNCG-GST fusion proteins were expressed in E.coli. The binding activities of anti-synuclein-gamma (SNCG) monoclonal antibodies (mAbs) with truncated SNCG were analyzed by ELISA and Western blot. According to the speculative epitope regions, expression vectors of potential epitopes were constructed and the GST-epitope fusion proteins were expressed in E.coli. And the fusion proteins were analyzed by ELISA and Western blot with mAbs to confirm the epitope regions.
Results: Epitope regions recognized by eleven anti-SNCG mAbs were identified. The epitopes recognized by 9#, 42#, and 3E4 were located in the region from 1st to 21st amino acids of SNCG, 11#, 22# in the region from 78th to 92nd amino acids, 1#, 14#, 18#, 31#, 36# in the region from 93rd to 110th amino acids, and 6C8 in the region from 111th to 127th amino acids.
Conclusion: The epitope regions recognized by eleven anti-SNCG mAbs were identified. It will benefit the research on SNCG and application of anti-SNCG monoclonal antibodies.